Apoptosis And Autophagy Of Cervical Cancer Cells Induced By Bufalin And Its Mechanism

Objective:Cervical cancer is one of the most common malignancies of the female reproductive system.Multiple drug resistance(MDR)induced by the tolerance of tumor cells to apoptosis is a major problem in the treatment of advanced cervical cancer.Therefore,it is important to develop new drugs to induce cervical cancer cell death by non-apoptotic pathways.Bufalin(bufalin)is an important active ingredient extracted from the skin and posterior ear gland of toad.A large amount of research evidence shows that bufalin can inhibit the growth of breast cancer,stomach cancer,lung cancer,bowel cancer and other tumors.But there has been no report on the treatment of cervical cancer.Autophagy is an important defense and protection mechanism of cells under stress.It can package and degrade damaged macromolecular proteins or organelles,produce new energy substances for cell recycling,and thus maintain homeostasis of the intracellular environment,and is also a way to induce tumor cell death.In our previous work,we found that C33 A cells and HeLa cells of cervical cancer treated with bufalin showed morphological changes of autophagy through electron microscopy.The purpose of this study was to explore the molecular mechanism of autophagy induced by bufalin in cervical cancer cells,so as to provide a theoretical basis for the clinical application of bufalin in the treatment of cervical cancer.Methods: 1.The killing effect of bufalin on cervical cancer cells C33 A and HeLa was detected by cck-8 method.2.Flow cytometry was used to detect the apoptosis induced by bufalin in cervical cancer cell lines.3.The pan-caspase inhibitor z-vad-fmk reverses the killing effect of bufalin on cervical cancer cells.4.Transmission electron microscopy was used to observe the effect of bufalin on autophagosome formation in cervical cancer cells.5.ROS levels of cervical cancer cells were detected by flow cytometry dcfh-da probe staining.6.Western blot method determination of bufalin LC3-?/? protein and JNK,p-JNK protein expression.7.The effect of ROS inhibitor NAC and JNK inhibitor SP600125 on the expression of LC3-? / ? protein in cervical cancer cells treated with bufalin.Results: 1.The killing effect of Bufalin on cervical cancer cells C33 A and HeLa was dose-time dependent.2.Bufalin induced apoptosis and autophagy of cervical cancer cell lines.3.Bufalin increases the production of ROS in cervical cancer cell lines.4.Bufalinactivates the JNK signaling pathway.5.NAC ROS inhibitors and JNK inhibitors SP600125 can be reversed bufalin caused by increase of LC3-? expression.Conclusion: Bufalin can induce apoptosis and autophagy of cervical cancer cells C33 A and HeLa,killing cervical cancer cell lines;Bufalin regulates cervical cancer cell autophagy through ROS / JNK signaling pathway.